Description
Broad Linear Range and Specific hKi-67 Detection
The Lumit® Cell Proliferation Assay demonstrates excellent linearity in microplate formats and specifically detects hKi-67 levels.
Measure Decreases in Cell Proliferation
Treatment of Jurkat cells (10,000/well, 384-well plate, ¼ standard 96-well volumes) with increasing concentrations of compounds. Both compounds reduced hKi-67 expression levels in a dose-dependent manner; however, BAY-1895344 induced cytotoxicity. Palbociclib produced a large change in hKi-67 levels without causing cytotoxicity after only 24 hours of treatment, enabling earlier assessment of compound effects on proliferation. Palbociclib’s effect appears minimal based on total ATP content.
Measure Increases in Cell Proliferation
Human CD8+ T cells (StemCell Tech) plated at 80,000 cells per well were treated with CD3/CD28 T cell activator with ( teal ) and without ( purple ) 10ng/ml IL-2 for 48 hours. Upregulation of hKi-67 is observed with the Lumit® Cell Proliferation Assay before T cell proliferation (which begins >72 hours after activation), demonstrating use of this assay as an early indicator of proliferation.
Compatible with 3D Culture Methods
In 3D spheroids, decreases in hKi-67 protein levels provide a clear and early indication of antiproliferative activity with a larger response window than a metabolic measure of viable cell number. HCT116 cells (1,000 cells/well) were plated in SBio PrimeSurface 96U plates (ULA; round bottom) and grown for 3 days to form 3D spheroids. The resultant HCT116 spheroids were then treated for 24 hours with increasing concentrations of nutlin-3a. Subsequently, proliferation was assessed with the Lumit® Cell Proliferation Assay (Human Ki-67) and a metabolic activity assay in separate plates. Note: Day 4 untreated HCT116 spheroids were ~440µm in diameter.
